The present invention relates to an aqueous solution as an eluent used in liquid chromatography for analysis of a specimen obtained from a human living body, comprising a mixture of acetonitrile and an aqueous solution of a member selected from the group consisting of acetic acid, a polybasic organic acid, a salt of organic acid and ammonium carbonate, and to a method for analyzing a specimen obtained from a human living body, comprising subjecting the specimen obtained therefrom to liquid chromatography while using, as an eluent, a mixture of acetonitrile and an aqueous solution of a member selected from acetic acid, a polybasic organic acid, a salt of organic acid and ammonium carbonate. More particularly, the present invention relates to an aqueous solution as eluent used in liquid chromatography for analysis of a specimen obtained from a human living body in order to detect a specific peak or a specific peak pattern intercorrelating to the hepatic morbid state, comprising a mixture of acetonitrile and an aqueous solution of a member selected from the group consisting of acetic acid, a polybasic organic acid, a salt of organic acid and ammonium carbonate, the volume ratio of acetonitrile to said aqueous solution of acetic acid or ammonium carbonate being 5:95 to 25:75, preferably 5:95 to 15:85, and the volume ratio of acetonitrile to said aqueous solution of the polybasic organic acid or the salt of organic acid being 1:99 to 25:75, preferably 5:95 to 15:85, and to a method for analysing a specimen obtained from a human living body in order to detecting a specific peak or specific peak pattern intercorrelating to the hepatic morbid state, comprising subjecting the specimen obtained therefrom to liquid chromatography while using, as an eluent, a mixture of acetonitrile and an aqueous solution of a member selected from acetic acid, a polybasic organic acid, a salt of organic acid and ammonium carbonate, the volume ratio of acetonitrile to said aqueous solution of acetic acid or ammonium carbonate is 5:95 to 25:75, preferably 5:95 to 15:85, and the volume ratio of acetonitrile to said aqueous solution of the polybasic organic acid or the salt of organic acid is 1:99 to 25:75, preferably 5:95 to 15:85.
It is extremely important for the diagnosis and treatment of a disease of a patient to analyze the components and the properties of a specimen such as blood, plasma, serum, cerebrospinal fluid, urine etc. taken from the patient and to obtain the information(s) concerning the morbid state of the patient based on the thus obtained analytical data. Such informations have been hitherto obtained by subjecting such a specimen to various chemical or biochemical analytical means, however, for the purpose of elucidating the various morbid states in various cases or seizing the more exact morbid state, the development of analytical methods by which more accurate informations are available has been demanded.
Particularly, in the cases of hepatic diseases and nephrotic diseases, because of the complicated morbid states thereof, the analytical methods which can provide new indices closely related to the morbid states of hepatic diseases and nephrotic diseases have been demanded.
As the conventional index of the morbid states of the hepatic diseases, the activity values of enzymes in the living body, for instance, GOT(glutamic-oxaloacetic transaminase), GPT(glutamic-pyruvic transaminase), LDH (lactic acid dehydrogenase), LAP(leucine aminopeptidase), etc., the biochemical analytical values of the blood components such as protein, lipoprotein, neutral fat, bilirubin, cholesterol etc. may be used.
However, it is difficult to trace precisely the change of the morbid state of a hepatic disease which causes the generation of very complicated factors in the patient suffering from the hepatic disease only by the utilization of the values obtained by the chemical analyses and biochemical analyses.
Accordingly, a new trial for diagnosing the morbid state of hepatic diseases while utilizing antigen-antibody reaction has been carried out (refer to Gastr., 76, 665(1979)), and it was reported that chronic active hepatitis and chronic inactive hepatitis or liver cirrhosis can be diagnosed at the respective probabilities of 50 to 60% and 20 to 40%, however, the method is complicated in its operations and is not satisfactory from the viewpoint of the accuracy in judging.
On the other hand, utilization of liquid chromatography is now attracting attention. Liquid chromatography is one of the analytical techniques based on the principles different from those on which the chemical or biochemical analytical method stands.
Utilization of liquid chromatography in the medical and clinical fields has been greatly expected in principle from the viewpoints that even the thermally and/or chemically unstable substances can be isolated and detected by liquid chromatography without being denaturated and that liquid chromatography is able to analyse many components in one operation while using a relatively minute amount of specimen, and a number of trials have been carried out in utilizing liquid chromatography.
However, in general, the peaks or the peak pattern in the chromatogram obtained by liquid chromatography are different to each other case by case where the eluents are different, and it changes according to the mode of combination of the eluent and the specimen, and accordingly, it is difficult to seize the peak, which directly correlates to the morbid state in a chromatogram. Particularly, in hepatic diseases, a method by which the morbid state of each of the various hepatic diseases including chronic hepatitis, liver cirrhosis etc. can be traced has not been established.
As a result of the present inventor's studies for the object of finding a method of clinical analysis of the specimen obtained from the patient suffering from a hepatic disease, in which an extremely small amount of the specimen is subjected to liquid chromatography, thereby obtaining a fraction exhibiting a peak which correlates to the progress of morbid state or the degree of seriousness of the disease within a short time period in a simple manner and quantifying the peak to examine the change of the morbid state from the change of the thus quantified peak appearing in the fractions, the present inventor has found a method fulfilling the object, and has attained the present invention.